Common housekeeping proteins are upregulated in colorectal adenocarcinoma and hepatocellular carcinoma, making the total protein a better "housekeeper"
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Xiaowen Hu1,*, Shujiao Du1,*, Jiekai Yu2, Xuhan Yang1, Chao Yang1, Daizhan Zhou1, Qingyu Wang1, Shengying Qin1, Xiaomei Yan3, Lin He1, Dongmei Han4, Chunling Wan1
1Bio-X Institutes, Key Laboratory for The Genetics of Developmental and Neuropsychiatric Disorders, Ministry of Education, Shanghai Jiao Tong University, Shanghai, 200030, PR China
2Cancer Institute, Key Laboratory of Cancer Prevention and Intervention, The Second Affiliated Hospital, Zhejiang University, College of Medicine, Hangzhou, Zhejiang, 310009, PR China
3School of Life Science and Biotechnology, Shanghai Jiao Tong University, Shanghai, 200240, PR China
4Instrumental Analysis Center, Shanghai Jiao Tong University, Shanghai, 200240, PR China
*These authors contributed equally to this work
Chunling Wan, email: [email protected]
Dongmei Han, email: [email protected]
Keywords: normalization, endogenous control, housekeeping protein, total protein amount, tumor
Received: March 03, 2016 Accepted: August 09, 2016 Published: August 20, 2016
Housekeeping proteins are essential endogenous controls for normalization as they are expected to be stably expressed. However, the stability of the expression level of housekeeping proteins needs to be assessed considering various experimental conditions. Our study evaluated the degree of variability of 7 commonly used housekeeping proteins with regard to their potential utility as normalizers in 56 pairs of matched colorectal adenocarcinoma (CRC) tissue samples and 6 pairs of hepatocellular carcinoma (HCC) tissue samples using multiple reaction monitoring (MRM) and Western blot analyses. A comprehensive experimental design and strict statistical analysis revealed that the expression levels of these 7 housekeeping proteins were not as stable as expected and they all exhibited upregulations to varying degrees in both the CRC and the HCC tissue samples. Consequently, we verified that using the amount of total protein instead of that of an individual protein can serve as a preferable control for studies of protein expression that require normalization.
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